The N-Terminal Domain and Glycosomal Localization of Leishmania Initial Acyltransferase LmDAT Are Important for Lipophosphoglycan Synthesis

Al-Ani, Gada; Patel, Nipul; Pirani, Karim A.; Zhu, Tongtong; Dhalladoo, Subbhalakshmi; Zufferey, Rachel

doi:10.1371/journal.pone.0027802

Cited by 4 publications

(6 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…To assign protein enrichment across the subcellular fractions, a spectral count methodology was employed because it gave a more robust agreement with the known subcellular localization of the above training set of 161 soluble proteins. Examination of the glycosomal markers (dihydroxyacetone phosphate acyltransferase, hexokinase, GMP reductase, and IMPDH components of the ether phospholipid biosynthesis, glycolysis, and purine salvage pathways ,, ) as well as the membrane proteins peroxin (PEX) 2, 11, 12, and 13, and the transporter GAT2 − showed that these proteins were found primarily in fractions 27–31 (glycosomes) (Table and Table S2). Interestingly, other well-characterized glycosomal enzymes involved in purine salvage xanthine phosphoribosyltransferase (XPRT) and HGPRT , and the glycolytic enzyme triose phosphate isomerase were also detected in fractions 1–4 (with a density of ∼1.10 to 1.12 g/mL) of the glycosome fractionation (Table and Table S2).…”

Section: Resultsmentioning

confidence: 99%

Proteomic Profiling of Leishmania donovani Promastigote Subcellular Organelles

et al. 2018

View full text Add to dashboard Cite

To facilitate a greater understanding of the biological processes in the medically important Leishmania donovani parasite, a combination of differential and density-gradient ultracentrifugation techniques were used to achieve a comprehensive subcellular fractionation of the promastigote stage. An in-depth label-free proteomic LC-MS/MS analysis of the density gradients resulted in the identification of ∼50% of the Leishmania proteome (3883 proteins detected), which included ∼645 integral membrane proteins and 1737 uncharacterized proteins. Clustering and subcellular localization of proteins was based on a subset of training Leishmania proteins with known subcellular localizations that had been determined using biochemical, confocal microscopy, or immunoelectron microscopy approaches. This subcellular map will be a valuable resource that will help dissect the cell biology and metabolic processes associated with specific organelles of Leishmania and related kinetoplastids.

show abstract

Section: Resultsmentioning

confidence: 99%

Proteomic Profiling of Leishmania donovani Promastigote Subcellular Organelles

et al. 2018

View full text Add to dashboard Cite

show abstract

“…DHAPAT enzymes from Leishmania ( Al-Ani et al, 2011 ) and Trypanosoma ( Zufferey et al, 2017 ) are much larger than their mammalian counterparts because they contain an N-terminal extension of unknown function. Interestingly, the ciliate Tetrahymena encodes the same bifunctional FARAT protein ( Dittrich-Domergue et al, 2014 ) as Dictyostelium , where the N-terminal extension encodes a functional fatty acid reductase domain ( Dittrich-Domergue et al, 2014 ) that may provide the fatty alcohol for the subsequent step in ether lipid synthesis ( Fig.…”

Section: Discussionmentioning

confidence: 99%

Phospholipids containing ether-bound hydrocarbon-chains are essential for efficient phagocytosis and neutral lipids of the ester-type perturb development in Dictyostelium

Kappelt

Hasna

et al. 2020

Biology Open

View full text Add to dashboard Cite

Lipids are the building blocks for cellular membranes; they provide signalling molecules for membrane dynamics and serve as energy stores. One path of their synthesis is initiated by glycerol-3-phosphate acyltransferase (GPAT), which in Dictyostelium resides on the endoplasmic reticulum. When an excess of fatty acids is present, it redistributes to storage organelles, the lipid droplets. Mutants, where the GPAT was eliminated by homologous recombination, produce fewer lipid droplets and are almost devoid of triacylglycerols (TAG), rendering them more resistant to cell death and cell loss in the developmental stages preceding fruiting body formation. The enzyme most closely related to GPAT is called FARAT, because it combines a fatty acyl-reductase (FAR) and an acyltransferase (AT) domain in its sequence. The protein is confined to the lumen of the peroxisome, where it transfers a fatty acid to dihydroxyacetone-phosphate initiating the synthesis of ether lipids, later completed at the endoplasmic reticulum. A mutant lacking FARAT produces lipid droplets that are devoid of the storage lipid monoalkyl-diacyl-glycerol (MDG), but the efficiency of spore formation in the developmental cycle is largely unaltered. Instead, these mutants are strongly impaired in phagocytosis of yeast particles, which is attributed to reduced synthesis of membrane phospholipids containing ether-linked chains.

show abstract

“…Additionally, glycolysis compartmentalization in the glycosomes prevents the accumulation of toxic intermediates generated during glucose degradation [35] . A recent study showed that glycosomal localization of Leishmania major dihydroxyacetonephosphate acyltransferase is important for lipophosphoglycan synthesis [36] .…”

Section: Discussionmentioning

confidence: 99%

Leishmania amazonensis Arginase Compartmentalization in the Glycosome Is Important for Parasite Infectivity

et al. 2012

View full text Add to dashboard Cite

In Leishmania, de novo polyamine synthesis is initiated by the cleavage of L-arginine to urea and L-ornithine by the action of arginase (ARG, E.C. 3.5.3.1). Previous studies in L. major and L. mexicana showed that ARG is essential for in vitro growth in the absence of polyamines and needed for full infectivity in animal infections. The ARG protein is normally found within the parasite glycosome, and here we examined whether this localization is required for survival and infectivity. First, the localization of L. amazonensis ARG in the glycosome was confirmed in both the promastigote and amastigote stages. As in other species, arg− L. amazonensis required putrescine for growth and presented an attenuated infectivity. Restoration of a wild type ARG to the arg − mutant restored ARG expression, growth and infectivity. In contrast, restoration of a cytosol-targeted ARG lacking the glycosomal SKL targeting sequence (argΔSKL) restored growth but failed to restore infectivity. Further study showed that the ARGΔSKL protein was found in the cytosol as expected, but at very low levels. Our results indicate that the proper compartmentalization of L. amazonensis arginase in the glycosome is important for enzyme activity and optimal infectivity. Our conjecture is that parasite arginase participates in a complex equilibrium that defines the fate of L-arginine and that its proper subcellular location may be essential for this physiological orchestration.

show abstract

The N-Terminal Domain and Glycosomal Localization of Leishmania Initial Acyltransferase LmDAT Are Important for Lipophosphoglycan Synthesis

Cited by 4 publications

References 34 publications

Proteomic Profiling of Leishmania donovani Promastigote Subcellular Organelles

Proteomic Profiling of Leishmania donovani Promastigote Subcellular Organelles

Phospholipids containing ether-bound hydrocarbon-chains are essential for efficient phagocytosis and neutral lipids of the ester-type perturb development in Dictyostelium

Leishmania amazonensis Arginase Compartmentalization in the Glycosome Is Important for Parasite Infectivity

Contact Info

Product

Resources

About